Fig 1: Effect of emodin on inflammatory factors and the FXR signaling pathway in ANIT-induced CLI mice. (A) The mRNA expression of inflammatory factors (IL-6, IL-1β, and TNFα) was determined by qRT-PCR. N = 5. (B) The mRNA expression of FXR, SHP, NTCP, BSEP, and MRP2 was determined using qRT-PCR. N = 5. (C,D) Representative WB images and quantitative analysis of FXR, SHP, BSEP, and NTCP protein expression in liver tissues. N = 3 independent biological replicates per group. (E) Representative immunofluorescence images of BSEP and NTCP (red) in the liver sections. Nuclei are counterstained with DAPI (blue). All data are presented as mean ± SD. *P < 0.05 vs. Control, #P < 0.05 vs. Model, ns P > 0.05 vs. Model.
Fig 2: Impact of FXR inhibition on emodin-mediated hepatoprotection and FXR pathway activation in ANIT-induced CLI mice (A) Representative images of HE-stained liver sections. (B) Serum levels of ALT, AST, TBA, and DBIL. N = 8. (C) Hepatic mRNA expression of inflammatory cytokines (IL-1β, IL-6, and TNF-α) was quantified by qRT-PCR. N = 3. (D) The mRNA expression of FXR and its target genes (SHP, BSEP, NTCP, and MRP2) was quantified by qRT-PCR. N = 3. (E,F) Representative WB images and quantitative analysis of FXR, SHP, BSEP, and NTCP protein expression in liver tissues. N = 3 independent biological replicates per group. Data are presented as the mean ± SD. *P < 0.05 vs. Control; #P < 0.05 vs. Model; †P < 0.05 vs. Emodin.
Fig 3: Effect of physcion on inflammatory factors and the FXR signaling pathway in ANIT-induced CLI mice. (A) The expression of inflammatory factors (IL-6, IL-1β, and TNFα) was determined by qRT-PCR. N = 5. (B) The expression of FXR, SHP, NTCP, BSEP, and MRP2 was determined using qRT-PCR. N = 5. (C,D) Representative WB images and quantitative analysis of FXR, SHP, BSEP, and NTCP protein expression in liver tissues. N = 3 independent biological replicates per group. (E) Representative immunofluorescence images of BSEP and NTCP (red) in the liver sections. Nuclei are counterstained with DAPI (blue). All data are presented as mean ± SD. *P < 0.05 vs. Control, #P < 0.05 vs. Model, ns P > 0.05 vs. Model.
Fig 4: Impact of FXR inhibition on physcion-mediated hepatoprotection and FXR pathway activation in ANIT-induced CLI mice (A) Representative images of HE-stained liver sections. (B) Serum levels of ALT, AST, TBA, and DBIL. N = 8. (C) Hepatic mRNA expression of inflammatory cytokines (IL-1β, IL-6, and TNF-α) was quantified by qRT-PCR. N = 3. (D) The mRNA expression of FXR and its target genes (SHP, BSEP, NTCP, MRP2) was quantified by qRT-PCR. N = 3. (E,F) Representative WB images and quantitative analysis of FXR, SHP, BSEP, and NTCP protein expression in liver tissues. N = 3 independent biological replicates per group. Data are presented as the mean ± SD. *P < 0.05 vs. Control; #P < 0.05 vs. Model; †P < 0.05 vs. Physcion.
Fig 5: Binding of emodin and physcion to FXR. (A–C) Predicted binding modes of GW4064 (A, slate), emodin (B, yellow), and physcion (C, orange) in the FXR (cyan) binding pocket. (a) 2D binding mode. (b,c) 3D binding mode. (D,E) FAC for determining the KD of emodin (D) and physcion (E) on FXR columns: breakthrough curves and nonlinear fitting curves. (F–H) SPR analysis of GW4064 (F), emodin (G), and physcion (H).
Supplier Page from Sino Biological, Inc. for Human NR1H4 Protein (His Tag)